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Using Nucleases to Stimulate Homologous Recombination

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Genetic Recombination

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 262))

Abstract

In essentially all organisms, double-strand breaks in chromosomal DNA stimulate repair by multiple mechanisms, including homologous recombination. It is possible to use site-specific reagents to produce a break or other recombinagenic damage at a unique site, which makes possible detailed analysis of the repair products. In addition, targeted mutagenesis and gene replacement are stimulated in the immediate vicinity of the break site. To utilize meganucleases with long recognition sequences, it is necessary to introduce the corresponding sequence prior to directed cleavage. The same is typically true of triplex-forming oligonucleotides that target polypurine-polypyrimidine tracts. Zinc-finger nucleases have the potential of being targetable to arbitrarily selected sites, owing to the flexibility of zinc finger recognition of DNA.

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Author information

Authors and Affiliations

  1. Department of Biochemistry II, University of Utah School of Medicine, Salt Lake City, UT

    Dana Carroll

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  1. Dana Carroll
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Editors and Affiliations

  1. Department of Biological Sciences, University of South Carolina, Coumbia, SC

    Alan S. Waldman

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© 2004 Humana Press Inc.

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Carroll, D. (2004). Using Nucleases to Stimulate Homologous Recombination. In: Waldman, A.S. (eds) Genetic Recombination. Methods in Molecular Biology™, vol 262. Humana Press. https://doi.org/10.1385/1-59259-761-0:195

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  • DOI: https://doi.org/10.1385/1-59259-761-0:195

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-236-0

  • Online ISBN: 978-1-59259-761-1

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